Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish

Short hairpin RNA (shRNA) directed by RNA polymerase III (Pol III) or Pol II promoter was shown to be capable of silencing gene expression, which should permit analyses of gene functions or as a potential therapeutic tool. However, the inhibitory effect of shRNA remains problematic in fish. We demonstrated that silencing efficiency by shRNA produced from the hybrid construct composed of the CMV enhancer or entire CMV promoter placed immediately upstream of a U6 promoter. When tested the exogenous gene, silencing of an enhanced green fluorescent protein (EGFP) target gene was 89.18 +/- 5.06% for CMVE-U6 promoter group and 88.26 +/- 6.46% for CMV-U6 promoter group. To test the hybrid promoters driving shRNA efficiency against an endogenous gene, we used shRNA against no tail (NTL) gene. When vectorized in the zebrafish, the hybrid constructs strongly repressed NTL gene expression. The NTL phenotype occupied 52.09 +/- 3.06% and 51.56 +/- 3.68% for CMVE-U6 promoter and CMV-U6 promoter groups, respectively. The NTL gene expression reduced 82.17 +/- 2.96% for CMVE-U6 promoter group and 83.06 +/- 2.38% for CMV-U6 promoter group. We concluded that the CMV enhancer or entire CMV promoter locating upstream of the U6-promoter could significantly improve inhibitory effect induced by the shRNA for both exogenous and endogenous genes compared with the CMV promoter or U6 promoter alone. In contrast, the two hybrid promoter constructs had similar effects on driving shRNA.

A 5.6-mW Power Dissipation CMOS Frequency Synthesizer for L1/L2 Dual-Band GPS Application

This paper presents a wide tuning range CMOS frequency synthesizer for dual-band GPS receiver, which has been fabricated in a standard 0.18-um RF CMOS process. With a high Q on-chip inductor, the wide-band VCO shows a tuning range from 2 to 3.6GHz to cover 2.45GHz and 3.14GHz in case of process corner or temperature variation, with a current consumption varying accordingly from 0.8mA to 0.4mA, from a 1.8V supply voltage. The measurement results show that the whole frequency synthesizer costs a very low power consumption of 5.6mW working at L I band with in-band phase noise less than -82dBc/Hz and out-of-band phase noise about -112 dBc/Hz at 1MHz offset from a 3.142GHz carrier.

Robust Tensor Analysis With L1-Norm

Tensor analysis plays an important role in modern image and vision computing problems. Most of the existing tensor analysis approaches are based on the Frobenius norm, which makes them sensitive to outliers. In this paper, we propose L1-norm-based tensor analysis (TPCA-L1), which is robust to outliers. Experimental results upon face and other datasets demonstrate the advantages of the proposed approach.

L1-Norm-Based 2DPCA

In this paper, we first present a simple but effective L1-norm-based two-dimensional principal component analysis (2DPCA). Traditional L2-norm-based least squares criterion is sensitive to outliers, while the newly proposed L1-norm 2DPCA is robust. Experimental results demonstrate its advantages.

重庆西山坪西园建材厂滑坡应急抢险治理工程设计与研究

以西山坪西园建材厂滑坡体工程地质勘察资料为依据,分析了滑坡体的岩土组成和物理力学性质以及造成滑坡的原因.认为影响山体滑坡的因素除与边坡地层岩性、微地貌特征、气象及水文条件有关外,修建厂房开挖与切割边坡坡脚则是重要的诱发原因.定性得出该滑坡的形成机制和类型为牵引式、浅层、小型土质新型滑坡.在此基础上,应用规范所推荐的极限平衡法,对该滑坡体的3个主要地质剖面(A-A'、B-B'、C-C')进行了稳定性计算与分析,得到不同工况下滑坡体稳定系数值.其中,在最不利组合工况(自重+暴雨+地震)下,滑坡体稳定系数仅为0.99～1.03,处于不稳定状态.根据计算结果,按照<长江三峡库区滑坡灾害防治规范>要求及"技术可行、安全可靠、经济合理、简单易实施"的设计原则,对该滑坡的应急抢险方案进行了设计,提出以抗滑桩拦挡工程为主、坡顶修筑截排水沟、地表裂缝回填夯实等综合治理方案.抗滑桩设置1排,分3段,根据每段不同的剩余下滑力设计抗滑桩的断面尺寸和埋设深度,使该滑坡体治理后,其安全系数分别满足不同工况下的规范要求.该方案已通过了三峡库区滑坡灾害防治专家组的审核,目前,正处于实施阶段.

Effects of rainfall infiltration on deep slope failure

With the finite element method and the limit equilibrium method, a numerical model has been estab-lished for examining the effects of rainfall infiltration on the stability of slopes. This model is able to reflect the variations in pore water pressure field in slopes, dead weight of the soil, and soil softening caused by rainfall infiltration. As a case study, an actual landslide located at the Nongji Jixiao in Chongqing was studied to analyze the effects of rainfall infiltration on the seepage field and slope sta-bility. The simulated results showed that a deep slope failure is prone to occur when rainfall infiltration leads to a remarkable variation in the seepage field, especially when the pore water pressure in slopes increases in a large range.

高桩码头不均匀沉降模型实验与数值模拟

以使用5年以上的高桩码头的大管桩为研究对象,采用模型实验的方法对大管桩的不均匀沉降进行了系统研究.首先通过量纲分析,确定模型实验的各个物理量和相关参数,以及模型材料(PVC管)等;然后利用电测技术,经过大量实验,获得大管桩的不均匀沉降量和变化规律;同时,利用数值模拟,分析了大管桩不均匀沉降变化规律,验证了模型实验结果的可靠性,说明了利用数值模拟方法分析大管桩不均匀沉降是可行的.

Effect of oxygen substitution by halide ions on structure, thermal stability and upconversion fluorescence in Er3(+)/Yb3(+)-codoped germanium-bismuth glasses

For the first time. effect of halide ions (F-, Cl-, Br-, and I-) introduction on structure, thermal stability, and upconversion fluorescence in Er3+/Yb3+-codoped oxide-halide germanium-bismuth glasses has been systematically investigated. The results show that halide ions modified germanium-bismuth glasses have lower maximum phonon energy and phonon density, worse thermal stability. longer measured lifetimes of I-4(l1/2) level, and stronger upconversion emission than germanium-bismuth glass. All these results indicate that halide ions play an important role in the formation of glass network, and have an important influence on the upconversion luminescence. The possible upconversion mechanisms of Er3+ ion are also evaluated. © 2005 Elsevier Ltd. All rights reserved.

不同氮肥管理措施对草坪生长和无机氮淋洗的影响

本研究采用室外盆栽试验，模拟运动场坪床结构，研究不同氮肥种类、不同施氮频率、施氮和降雨时间间隔对草地早熟禾草坪质量、草坪生长和无机氮淋洗的影响，并探讨在北京地区气候条件下，草坪在不同氮肥管理措施下的氮素去向及环境风险。主要结论如下： 1. 氮肥种类对草坪质量和草坪生长有显著影响。试验前期（春季），草坪颜色和密度质量、草坪草生长速度和草屑全氮含量的排序为尿素 > CU3M（自研包膜尿素）> IBDU（进口缓释肥），而试验后期（秋季）则为CU3M = IBDU > 尿素。草坪合格颜色质量持续时间和成坪速度的排序为尿素 > CU3M > IBDU。在新建草坪选择缓释肥进行早春施肥时，应混施一定比例的速效肥。 2. 施氮频率对草坪外观质量评分、草坪生长速度、草屑总生物量、草屑全氮含量和根系分布状况有显著影响。U6处理的草坪合格颜色质量持续时间最长，增加施氮频率没有降低新建草坪生长速度和草屑全氮含量的波动幅度。 3. 夏季（6~8月）渗漏液体积占全年的比例最高，为75.0%~82.4%。新建草坪初期的淋洗风险较大，渗漏液硝态氮浓度在第1次超过10 mg N•L-1，尿素分6次施用可降低这次的渗漏液硝态氮浓度。在合理施氮量内，草坪成熟后的无机氮淋洗量很小，且草坪对雨水中的无机氮有吸收和过滤作用。 4. 春季施用氮肥的吸收利用率为48.0%~72.6%，草屑吸收量最高，占38.5%~48.7%，地上部吸收量占19.6%~22.1%，根系吸收量在7.3%以下。施肥处理的无机氮淋洗损失量仅为0.23~0.42 g N•m-2，与CK无显著差异，草地早熟禾12 g N•m-2的年施氮量对环境的风险很小。 5. 初秋施氮5 g N•m-2，可以保持草地早熟禾秋季的良好颜色和密度质量，草坪草氮素吸收利用率高达87.4%~99.7%，其中草屑带走量占24.3%~34.2%，地上部吸收量占43.3%~59.6%，根系吸收量占14.2%~19.1%。 6. 施氮和模拟降雨的时间间隔对渗漏液硝态氮浓度有显著影响，间隔6 d和9 d模拟降雨后的渗漏液硝态氮浓度最高，显著高于间隔3 d和12 d模拟降雨的结果。初秋合理施氮的渗漏液硝态氮浓度在1.0 mg N•L-1 以下，环境风险较小。

家畜放牧对典型草原土壤-植被系统氮素转化的影响

草地生态系统中，放牧对调节物质流动和营养循环起着关键的作用。内蒙古地区已有上千年的游牧历史，放牧是该地区重要的草地利用方式之一。然而，近50年来，由于人口的剧增以及对草原的不合理利用与管理，使得内蒙古草原发生了严重的退化与沙漠化。理解放牧对氮循环的定量影响，对我们更合理地利用草地、防治生态系统的进一步退化以及探求最佳恢复途径都具有重要的意义。中国科学院内蒙古草原生态系统定位站精准设置的5种放牧强度处理（0.00, 1.33, 2.67, 4.00, 5.33 羊/公顷）为我们的研究提供了理想的平台。2005、2006年生长季期间，在经过16年不同放牧强度处理的一个典型草原样地上，我们测定了氮素输入（氮沉降、生物固氮）、转化（净氮矿化）、输出（反硝化及氨气挥发）速率等N循环的重要参量。同时，测定了微生物生物量碳、氮（Cmic，Nmic）及微生物呼吸（Rmic），研究了微生物在氮循环中的作用。另外，还测定了植物、土壤、固氮体中的15N自然丰度值，探讨了其对不同放牧强度的响应格局与机理。 结果表明，干湿混合沉降物全氮浓度最高达11.53 mg N l–1，沉降量最高为1.77 kg N ha–1m–1。月均氮沉降量与浓度正相关，它们与降水量均关系密切，前者更密切。结皮面积所占比例很小，不超过8%。结皮含氮量为1.01-1.43 g N kg–1，受放牧的影响不显著，但随放牧强度的增加有降低的趋势。土壤结皮能固氮，但固氮量不超过土壤氮含量的1倍。地耳的固氮量为结皮的10-20倍，故是主要的固氮体。尽管地耳含氮量受放牧影响不显著，但放牧是不放牧条件下地耳氮含量的1.58倍。 土壤NH4+-N浓度随季节变化范围为1.71-9.45 µg N g–1，它们在各放牧处理之间的差异不显著。土壤NO3–-N浓度变幅为0.27-11.21 µg N g–1。总无机氮浓度在不放牧条件下的变幅为2.69-14.57 µg N g–1，占总氮的0.49-2.6%;放牧条件下的变幅为2.49-8.66 µg N g–1，占总氮的0.35-1.21%。总无机氮浓度随季节和放牧强度的变化趋势与硝态氮相似，表现为2005年夏季期间有逐渐增加的趋势，而在2006年整个生长季期间有逐渐降低的趋势。不放牧比放牧条件下含氮量高，但在4个放牧处理之间的差异不显著。净氮矿化速率的变幅为–0.61-0.27 µg N g–1d–1，峰值通常出现在7月。净氮矿化速率在各处理间没有一致性差异，但中牧（2.67、4.00 羊/公顷）通常比重牧（5.33 羊/公顷）下的值高。净氮硝化速率通常很低，波动在–0.32-0.16 µg N g–1之间，2005年夏季及2006年春秋季的值相对较高。净氮硝化速率在各放牧处理之间差异不显著，但重牧条件下的值通常最低。累积净氮转化量在年际间差异大，2005年总体上遵循正态分布模式，而2006年随着放牧强度的增加有直线下降的趋势，2006年比2005年的累积量高。土壤温度和湿度比放牧强度对净氮矿化的影响更加显著。放牧强度通过调节这两个土壤因子对氮动态而产生间接影响。 反硝化和N2O的释放速率低，前者变幅为0.33-6.21µg N kg–1 d–1，后者为0.42-11.28 µg N kg–1 d–1。释放量夏季较强，春秋较弱。放牧对反硝化释放影响不显著，只在2005年对N2O释放影响显著。尽管如此，反硝化和N2O释放速率整体表现出在不放牧比放牧条件下高的趋势，且比最高放牧强度5.33 羊/公顷下的反硝化速率显著高。然而，它们在4个放牧处理之间的差异始终不显著。累积反硝化和N2O释放存在年际变化，2006年的值显著高。它们随着放牧强度的递增有逐渐降低的趋势，这在2006年表现得尤为明显，这种结果主要归因于土壤总氮量在长期放牧条件下随放牧压力的增加而逐渐降低。 氨气挥发速率变幅为0.88-3.52 g N ha–1d–1，高峰值出现在5月，2005比2006年同期的速率大。两年间放牧强度对氨气挥发的影响都较弱，2005年影响更弱。不放牧条件下的氨气挥发量通常最低，这在生长季的前期表现得更为明显，中牧及重牧条件下通常最高。放牧能影响氨气挥发与氨态氮，硝态氮及总无机氮浓度之间的关系，即不放牧条件下相关性显著，而放牧条件下相关性不显著。年际间氨气挥发速率与无机氮浓度之间的关系趋势相反，2005年负相关，2006年正相关。在水分充足的2006年，所有处理条件下氨气挥发与土壤水分及温度之间显著相关，但在单独每个放牧处理下，相关性不显著。 Cmic变幅大，为13.97-350.45 μg C g–1，占土壤总有机碳的1.58-8.35%。最高和最低值分别出现在夏季和春季。它们在各处理间差异不显著，不放牧下的值相对偏高。Cmic与土壤有机C和全N、前期的立枯、凋落量及含N量、优势种前期的地上生物量、根系生物量、土壤温度以及水分之间关系密切。氮素状态如氨态氮、总无机氮含量、反硝化以及N2O释放速率，氨气挥发速率与Cmic之间关系密切。Nmic占土壤全氮的0.41-2.74%，不受季节和放牧强度的显著影响。Nmic与可溶性N，表土层根系全N，立枯有机C，地上生物量之间关系密切。氮循环过程中氨气挥发速率受Nmic的影响。Rmic随季节而变化，通常5月份值最高。Rmic随着放牧强度的增加有稍降低的趋势。Rmic与土壤可溶性C、有机C，不同土层根系有机C，凋落物、立枯量及其C、N含量、全N，地上生物量，优势种前期的生物量，土壤温度之间关系密切。土壤氮循环动态如氨态氮、硝态氮、总无机氮浓度及反硝化速率与Rmic之间关系密切。 土壤、植物、地耳、生物结皮的δ15N值与放牧强度之间相关关系不显著。然而，放牧有增加表层土和植物的δ15N值而降低表土、地耳、结皮的δ15N值的趋势。表层土δ15N值与前一年生长季末期硝态氮及总无机氮浓度，反硝化速率及累积氨气挥发之间密切相关。 土壤碳含量的变幅为10.44-17.19 g C kg–1，全氮量的变幅为0.54-0.82 g N kg–1。长期的高强度放牧降低了土壤碳、氮储量。根系碳、氮含量分别为土壤碳、氮含量的40-50和10倍。立枯和凋落物有机碳含量变幅为446.94-507.01 g C kg–1，与放牧强度之间关系不密切;氮含量变幅为4.58-7.18 g N kg–1，与放牧强度之间显著负相关。优势种木地肤、冷蒿的含碳量与放牧强度之间相关不显著，但含氮量与之显著相关。 综述以上结果，不同放牧强度对内蒙草地生态系统氮循环中不同过程产生影响的程度各不相同，这种影响主要是通过它与土壤环境因子如温度、水分的联合作用而间接产生。

武汉月湖和莲花湖表层沉积物中持久性有机物的污染状况

采用气-质联用技术分析了武汉汉阳月湖和莲花湖的4个表层沉积物样品中的有机污染物,探讨了两湖沉积物受持久性有机物污染的程度。月湖中共检测出124种有机物,其中属环境优先控制污染物和美国EPA筛选的内分泌干扰物19种;莲花湖中共检测出186种有机物,环境优先控制污染物和美国EPA筛选的内分泌干扰物34种。主要污染物包括:酞酸酯、酯类、酚类、杂环和苯及其衍生物等。污染物浓度顺序为L1>L2>Y2>Y1,莲花湖中有机物浓度明显高于月湖。两湖邻苯二甲酸酯的含量最高,占了污染物总量的96%～98%,邻苯二甲酸乙基己基

Short and long peptidoglycan recognition proteins (PGRPs) in zebrafish, with findings of multiple PGRP homologs in teleost fish

Peptidoglycan recognition protein (PGRP) specifically binds to peptidoglycan and is considered to be one of the pattern recognition proteins in the innate immunity of insect and mammals. Using a database mining approach and RT-PCR, multiple peptidoglycan recognition protein (PGRP) like genes have been discovered in fish including zebrafish Danio rerio, Japanese pufferfish TakiFugu rubripes and spotted green pufferfish Tetraodon nigroviridis. They share the common features of those PGRPs in arthropod and mammals, by containing a conserved PGRP domain. Based on the predicted structures, the identified zebrafish PGRP homologs resemble short and long PGRP members in arthropod and mammals. The identified PGRP genes in T. nigroviridis and TakiFugu rubripes resemble the long PGRPs, and the short PGRP genes have not been found in T. nigroviridis and TakiFugu rubripes databases. Computer modelling of these molecules revealed the presence of three alpha-helices and five or six beta-strands in all fish PGRPs reported in the present study. The long PGRP in teleost fish have multiple alternatively spliced forms, and some of the identified spliced variants, e.g., tnPGRP-L3 and tnPGRP-L4 (in: Tetraodon nigroviridis), exhibited no characters present in the PGRP homologs domain. The coding regions of zfPGRP6 (zf: zebrafish), zfPGRP2-A, zfPGRP2-B and zfPGRP-L contain five exons and four introns; however, the other PGRP-like genes including zfPGRPSC1a, zfPGRPSC2, tnPGRP-L1-, tnPGRP-L2 and frPGRP-L (fr: Takifugu rubripes) contain four exons and three introns. In zebrafish, long and short PGRP genes identified are located in different chromosomes, and an unknown locus containing another long PGRP-like gene has also been found in zebrafish, demonstrating that multiple PGRP loci may be present in fish. In zebrafish, the constitutive expressions of zfPGRP-L, zfPGRP-6 and zfPGRP-SC during ontogeny from unfertilized eggs to larvae, in different organs of adult, and the inductive expression following stimulation by Flavobacterium columnare, were detected by real-time PCR, but the levels and patterns varied for different PGRP genes, implying that different short and long PGRPs may play different roles in innate immune response. (c) 2007 Elsevier Ltd. All rights reserved.

Purification and characterization of hydrosoluble components from the sap of Chinese lacquer tree Rhus vernicifera

Continuous gradient elution chromatography (CGEC) was employed to purify and separate enzymes and polysaccharides from the sap of Rhus vernicifera Chinese lacquer tree. There are three different molecules with laccase enzyme activity. Two are enzymes of each other (L1, and L2), whereas the third (RL) is an entirely separate entity. Two polysaccharides (GP1 and GP2) were also found. The Rhus laccase (RL), and isoenzymes L1 and L2, have peak molecular masses of 109,100, 120,000, 103,000 respectively; each has four copper atoms per molecule, and the pI values were 8.2, 8.6, and 9.1, respectively. The structure of the laccases was studied by Fourier-transform infrared (FT-IR) and Matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry. The typical amide I (1646 cm(-1)) and amide II (1545 cm(-1)) bands were observed. The results from MALDI-TOF were similar to those from CGEC, but the molecular mass from the MALDI-TOF was significantly different from that obtained from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). (c) 2006 Elsevier B.V. All rights reserved.

Characterization of cDNA encoding immunoglobulin light chain of the Mandarin fish (Siniperca chuatsi)

Immunoglobulin light chain cDNA sequences of a perciform fish, the mandarin fish Siniperca chuatsi were amplified from head kidney mRNA by reverse transcription (RT)-PCR and RACE methods using degenerated primer and gene specific ones. In cDNA sequences of the VL region, nucleotide exchanges were present mainly within CDRs, although a lesser degree of variability was also found in FRs. Moreover, the length of CDRI and CDR3 in the mandarin fish is shorter than in most other fish species. In the middle of S. chuatsi CL region, a microsatellite sequence (AGC)(6-8) was found, which is also present in another perciform species, the spotted wolffish (Anarhichas minor). The comparison of amino acid sequence of the mandarin fish CL domain with those of other vertebrates showed the highest degree of similarity of 94.5% to the spotted wolffish, while the similarity with rainbow trout (Oncorhynchus mykiss) Ig L1 (62.7%) and channel catfish (Ictalurus punctatus) Ig LG (55.9%) isotypes is also higher. However, there is only 50% identity in the VL regions between the mandarin fish and the wolffish. The sequence similarity of the mandarin fish CL domain with those of higher vertebrate did not readily allow it to be classified as kappa or lambda isotype. The phylogenetic analyses also demonstrated that the CL genes of the mandarin fish and most other teleost fish cluster as a separate branch out of the mammal kappa and lambda branches. (C) 2003 Elsevier B.V. All rights reserved.